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Figure S3. PCR of Necl-2 isoforms and Northern analysis of Necl-1. (A) PCR analysis of Necl-1 and -4 expression from first-strand templates generated from purified cultures of Schwann cells and DRG neuron enriched cultures are shown. Necl-4 and -1 amplified differentially from these templates. Some Schwann cells persist in the DRG neuron cultures, potentially accounting for the amplification of Necl-4 from these cultures. (B) The expression of six Necl-2 splice variants in Schwann cells and DRG neurons was analyzed by PCR. Both Schwann cells and DRG neurons express nearly the same complement of isoforms (1, 2, and 6) except that isoforms 3 and 4 are preferentially expressed by Schwann cells and DRG neurons, respectively. These splices variants encompass the mucin-like poly-threonine region, and, as such, isoform 3 yields an expected smear between 75 and 95 bp. Isoform 5 is not expressed at detectable levels in either Schwann cells or DRG neurons. (C) Tissue Northern blot analysis of Necl-1 demonstrates high level expression in central nervous system tissue and more modest expression in sciatic nerve. (D) Necl-1 mRNA is faintly detected during development of the sciatic nerve, with a peak at p10 and p15.